DESCRIPTION: State the application's broad, long-term objectives and specific aims, making reference to the health relatedness of the project. Describe concisely the research design and methods for achieving these goals. Avoid summaries of past accomplishments and the use of the first person. This abstract is meant to serve as a succinct and accurate description of the proposed work when separated from the application. If the application is funded, this description, as is, will become public information. Therefore, do not include proprietary/confidential information. DO NOT EXCEED THE SPACE PROVIDED. The goal of the project is to develop and demonstrate two related approaches to molecular analysis and separation that employ flow based analytical instrumentation and magnetic microsphere technology: a magnetic flow spectrometer for separation, and a magnetic flow cytometer for identification. The flow spectrometer system will be unique in enabling highly parallel continuous flow biomolecular separations on a preparative scale, streamlining downstream analysis and revolutionizing our ability to identify potential diagnostic or therapeutic targets. The magnetic flow cytometer will combine a novel magnetic target- molecule tagging concept with fluorescence-based analyte detection. The instrumentation proposed will contribute significantly to a broad range of applications improving human health and quality of life including drug discovery, molecular targeting, DNA analysis, proteomics, and understanding the pathways of cell cycle regulation. We will validate the new instrument by conventional molecular analysis methods and apply it to the study of intracellular vesicle traffic. A product for commercialization is anticipated. Operation of the proposed instrument involves three steps. 1) Magnetically encoded microspheres are prepared by encapsulating strong ferromagnetic material with high remnant magnetization and coercivity, never before used for such applications, in polymer spheres. The distribution of microspheres can be sorted into different bins depending on their intrinsic magnetic moment by flowing through a chamber where a magnetic field gradient induces a force such that they are collected in different bins with narrow distributions of magnetic moment. Microspheres from each bin are chemically bound to target molecules so that each species of magnetic moment is bound to one unique kind of molecule. The collection of microspheres and associated target molecules are then mixed together and incubated with analytes. 2) The incubated collection of microspheres are flowed through a SQUID detector system which identifies the target molecule by measuring the magnetic moment of the microsphere to which it is attached. 3) The analytes will be chemically prepared with molecular groups that fluoresce when illuminated by a laser beam, indicating the target-analyte binding. Combining SQUIDs for target identification with laser diagnostics to assess binding provides an efficient, high throughput multiplexed bioassay method based on traditional flow cytometry. PERFORMANCE SITE(S) (organization, city, state) Los Alamos National Laboratory Los Alamos NM 87545 University of Nebraska Lincoln Lincoln, NE. 68588 KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required information in the format shown below. Start with Principal Investigator. List all other key personnel in alphabetical order, last name first. Name Organization Role on Project Michelle Espy Physics Div., Biophysics & Quantum Inf. PI Robert Kraus Physics Div., Biophysics & Quantum Inf. Andrei Matlachov Physics Div., Biophysics & Quantum Inf. John Nolan Bioscience Div., Langham Resource Piotr Grodzinski Bioscience Div., Michelson Resource Diandra Leslie-Pelecky Physics Dept. University Nebraska, Lincoln Disclosure Permission Statement. Applicable to SBIRISTTR Only. See instructions. [] Yes [] No PHS 398 (Rev. 05/01) Page 2 Form Page 2 Principal Investigator/Program Director (Last, First, Middle): Espy MJchelle A The name of the principal investigator/program director must be provided at the top of each pdnted page and each continuation page. RESEARCH GRANT